Comparative Study on the Biological Characteristics of Japanese encephalitis virus Envelope Protein I176R Site Mutant Strain
TANG Mian-Yao1,2,3, ZHAO Qin1,2,3, SHI Yan-Ping1,2,3, CAO Yu-Qin1,2,3, WEN Yi-Ping1,2,3, WU Rui1,2,3, DU Sen-Yan1,2,3, HUANG Xiao-Bo1,2,3, YAN Qi-Gui1,2,3, CAO San-Jie1,2,3,*
1 College of Veterinary Medicine/Research Center for Swine Diseases, Sichuan Agricultural University, Chengdu 611130, China; 2 National Demonstration Center for Experimental Animal Education, Sichuan Agricultural University, Chengdu 611130, China; 3 Sichuan Science-Observation Experimental Station of Veterinary Drugs and Veterinary Diagnostic Technique, Ministry of Agriculture and Rural Affairs, Chengdu 61130, China
Abstract：Japanese encephalitis virus (JEV) is a serious zoonotic virus, and the envelope protein (E) plays a key role in the neurovirulence of JEV. Previous studies have found that the mutation of EI176R site of recombinant Japanese encephalitis virus (rJEV-EI176R) can weaken its neurovirulence. In order to identify the biological characteristics of rJEV-EI176R, the E gene sequence of rJEV-EI176R was cloned and analyzed, and the bioinformatics characteristics of E protein were analyzed. The growth curve of rJEV-EI176R, the adsorption difference of various cells and the inflammatory level of mouse microglia cell line (BV-2) induced by rJEV-EI176R were determined by qPCR, plaque assay and indirect immunofluorescence assay (IFA). The results showed that the positive charge residue of E protein of rJEV-EI176R increased by 1, the isoelectric point increased by 0.37, and the instability index increased by 0.39. The number and proportion of α-helix, β-sheet, extended strand and random coil of E protein changed, and the amino acid at E176 site had different connections with nearby amino acids. The growth curve of rJEV-EI176R on BV-2 cells did not change significantly and could infect nerve cells and renal cells from different sources. The adsorption capacity of rJEV-EI176R was significantly improved (P<0.05). After inoculation of BV-2 cells with rJEV-EI176R, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interferon-γ-induced protein-10 (IP-10) were significantly lower than those of the parental strain at 12 and 24 h (P<0.05). This study enriched the biological characteristics of E protein of JEV strain, and provides basic data for further study of JEV attenuated mechanism and vaccine development.
唐冕耀, 赵勤, 石艳萍, 曹玉琴, 文翼平, 伍锐, 杜森焱, 黄小波, 颜其贵, 曹三杰. 乙型脑炎病毒囊膜蛋白I176R位点突变株的生物学特性比较研究[J]. 农业生物技术学报, 2024, 32(10): 2391-2401.
TANG Mian-Yao, ZHAO Qin, SHI Yan-Ping, CAO Yu-Qin, WEN Yi-Ping, WU Rui, DU Sen-Yan, HUANG Xiao-Bo, YAN Qi-Gui, CAO San-Jie. Comparative Study on the Biological Characteristics of Japanese encephalitis virus Envelope Protein I176R Site Mutant Strain. 农业生物技术学报, 2024, 32(10): 2391-2401.
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